Paris/July 12
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Nicolas C. (Talk | contribs) |
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==Preparation of a stock of phage on w121:== | ==Preparation of a stock of phage on w121:== | ||
| + | David B: | ||
* Filtration of the Cacl2 and MgSO4 stocks | * Filtration of the Cacl2 and MgSO4 stocks | ||
* Launch a culture: | * Launch a culture: | ||
| Line 11: | Line 12: | ||
* Let grow until OD=0.2 (~1h) | * Let grow until OD=0.2 (~1h) | ||
* Add 100µl P1 (stock 10^10/ml) | * Add 100µl P1 (stock 10^10/ml) | ||
| - | |||
| - | |||
* Incubate 2h | * Incubate 2h | ||
* Add 200µl Chloroform | * Add 200µl Chloroform | ||
* Vortex, centrifuge (15min @ 3000rpm) | * Vortex, centrifuge (15min @ 3000rpm) | ||
| - | * Recover the supernatant = Stock keep at 4°C | + | * Recover the supernatant + add 200µl Chloroform = Stock keep at 4°C |
[[User:Nicolas C.|Nicolas C.]] | [[User:Nicolas C.|Nicolas C.]] | ||
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* The stock prepared the 8.7.7 | * The stock prepared the 8.7.7 | ||
See protocols for details. | See protocols for details. | ||
| + | |||
| + | == Culture of FtsZ TS == | ||
| + | We want to isolate a good clone of the strain FstZ TS | ||
| + | From the plate of FtsZ TS84 121: | ||
| + | isolation of 6 clones of the 121.1 on LBA+tet incubated at 42°C + Culture ON at 30°C | ||
Revision as of 14:00, 12 July 2007
Preparation of a stock of phage on w121:
David B:
- Filtration of the Cacl2 and MgSO4 stocks
- Launch a culture:
- 15ml LB
- 225µl Cacl2
- 450µl MgSO4
- 90µl DAP
- 150µl W121 from an ON culture
- Let grow until OD=0.2 (~1h)
- Add 100µl P1 (stock 10^10/ml)
- Incubate 2h
- Add 200µl Chloroform
- Vortex, centrifuge (15min @ 3000rpm)
- Recover the supernatant + add 200µl Chloroform = Stock keep at 4°C
Titration of bacteriophages P1
We measure the strength of former P1 preparation :
- The stock prepared the 3.7.7
- The stock prepared the 8.7.7
See protocols for details.
Culture of FtsZ TS
We want to isolate a good clone of the strain FstZ TS From the plate of FtsZ TS84 121: isolation of 6 clones of the 121.1 on LBA+tet incubated at 42°C + Culture ON at 30°C
